Live Tissue Intrinsic Emission Microscopy Using Multiphoton-Excited Native Fluorescence and Second Harmonic Generation

Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intri...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2003-06, Vol.100 (12), p.7075-7080
Main Authors: Zipfel, Warren R., Williams, Rebecca M., Christie, Richard, Nikitin, Alexander Yu, Hyman, Bradley T., Webb, Watt W.
Format: Article
Language:English
Subjects:
Alzheimer Disease - metabolism
Alzheimer Disease - pathology
Alzheimer's disease
Animals
Biological Sciences
Biophysical Phenomena
Biophysics
Cancer
Collagen - metabolism
Collagens
Emission spectra
Female
Fluorescence
Fluorescent Dyes
Histology
Humans
Imaging
Indoles - metabolism
Mammary Neoplasms, Experimental - metabolism
Mammary Neoplasms, Experimental - pathology
Mice
Microscopy
Microscopy, Fluorescence - methods
Molecular biology
NAD - metabolism
Neurofibrillary Tangles - metabolism
Neurofibrillary Tangles - pathology
Photons
Pollutant emissions
Solar fibrils
Tumors
Vitamin A - metabolism
Wavelengths
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Description
Summary:Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen provides crucial information for physiology and pathology. The efficient application of multiphoton microscopy to intrinsic imaging requires knowledge of the nonlinear optical properties of specific cell and tissue components. Here we compile and demonstrate applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0832308100